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- Axelsson Olsson, Diana, et al.
(author)
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A simple method for long-term storage Acanthamoeba species
- 2009
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In: Parasitology Research. - : Springer Science and Business Media LLC. - 0932-0113 .- 1432-1955. ; 104:4, s. 935-937
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Journal article (peer-reviewed)abstract
- We present a novel and simple technique for storing live Acanthamoeba for long periods of time. The amoebae are maintained at refrigerator temperatures in a peptone-yeast extract-glucose (PYG) medium normally used for cultivation. Using this method, we obtained survival rates of at least 4 years for Acanthamoeba polyphaga and 3 years for Acanthamoeba castellanii and Acanthamoeba rhysodes. Advantages of this storage method are: (1) it is quick and simple, (2) inexpensive, (3) does not require encystment before storage, (4) resuscitation of cysts can be achieved within a week of culture in PYG medium at 27A degrees C, and does not require co-culture with bacteria or any special equipment.
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| 2. |
- Axelsson Olsson, Diana, et al.
(author)
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Acanthamoeba-Campylobacter coculture as a novel method for enrichment of Campylobacter species
- 2007
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In: Applied and Environmental Microbiology. - 0099-2240 .- 1098-5336. ; 73:21, s. 6864-6869
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Journal article (peer-reviewed)abstract
- In this study, we present a novel method to isolate and enrich low concentrations of Campylobacter pathogens. This method, Acanthamoeba-Campylobacter coculture (ACC), is based on the intracellular survival and multiplication of Campylobacter species in the free-living protozoan Acanthamoeba polyphaga. Four of the Campylobacter species relevant to humans and livestock, Campylobacter jejuni, C. coli, C. lari, and C. hyointestinalis, were effectively enriched by the coculture method, with growth rates comparable to those observed in other Campylobacter enrichment media. Studying six strains of C. jejuni isolated from different sources, we found that all of the strains could be enriched from an inoculum of fewer than 10 bacteria. The sensitivity of the ACC method was not negatively affected by the use of Campylobacter-selective antibiotics in the culture medium, but these were effective in suppressing the growth of seven different bacterial species added at a concentration of 10(4) CFU/ml of each species as deliberate contamination. The ACC method has advantages over other enrichment methods as it is not dependent on a microaerobic milieu and does not require the use of blood or other oxygen-quenching agents. Our study found the ACC method to be a promising tool for the enrichment of Campylobacter species, particularly from water samples with low bacterial concentrations.
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| 3. |
- Axelsson Olsson, Diana, et al.
(author)
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Campylobacter jejuni acid tolerance increases when co-incubated with amoebae
- 2009
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Conference paper (peer-reviewed)abstract
- Background: Although Campylobacter jejuni is a frequent cause of bacterial gastroenteritis, one of the enigmas is how thisfragile organism can survive the transit through the acid milieu of the stomach. C. jejuni is very sensitive to low pH, but cansurvive in moderately acid environment for short periods of time. We have previously shown that C. jejuni can colonize andeven replicate in different species of amoebas, thereby gaining protection from adverse environments.Objectives: We evaluated the effects of hydrochloric acid (HCl) on C. jejuni at various pH and time intervals, to study whetherco-cultivation with amoeba influenced C.jejuni acid tolerance. The setup was chosen to mimic the acidified milieu of the humangastrointestinal tract.Methods: Cultures of C. jejuni (CCUG 11284) were co-cultured with Acanthamoeba polyphaga in either PBS or tap wateracidified with HCl to pH 1, 2, 3 and 4. We also evaluated different treatments effect on campylobacter survival, by exposingsome bacterial samples to an acid shock and some to a slower acidification process.Results and conclusions: We show that C. jejuni can withstand pH below the normal range of survival, when co-cultured withA. polyphaga. C. jejuni co-cultured with amoebae survived acidified conditions at pH 3 for 20 hours and pH 2 for approximately5 hours. We also found a pH increase during the experiment, which correlated with campylobacter survival. These results pointto an unknown mechanism for C.jejuni to survive at low pH levels. This could be in the form of excretion of pH-increasingsubstances and simultaneous chemotaxic orientation towards a protective host. Our results could give one possible explanationto C. jejuni survival through the low pH of the gastrointestinal tract.
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| 9. |
- Fischer, Hans, et al.
(author)
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Ceramide as a TLR4 agonist; a putative signalling intermediate between sphingolipid receptors for microbial ligands and TLR4.
- 2007
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In: Cellular Microbiology. - : Hindawi Limited. - 1462-5814 .- 1462-5822. ; 9:5, s. 1239-1251
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Journal article (peer-reviewed)abstract
- Mucosal Toll-like receptors (TLRs) respond to pathogens, but remain inert to the indigenous flora, suggesting that the TLRs can receive pathogen-specific signals. For example, TLR4 signalling is activated in CD14-negative epithelial cells by P-fimbriated, uropathogenic Escherichia coli, but not by lipopolysaccharide. The fimbriae use glycosphingolipids as recognition receptors and there is release of ceramide, which is the membrane-anchoring domain of the receptors. In this study, ceramide was identified as a TLR4 agonist and as a putative signalling intermediate between the glycosphingolipid recognition receptors and TLR4. Exogenous ceramide activated a TLR4-dependent epithelial cell response, as shown by exposing stably transfected TLR4-positive or -negative human embryonal kidney cells to C2 and C6 ceramide. A similar, TLR4-dependent response occurred after deliberate release of endogenous long-chained ceramide with sphingomyelinase. Microbial ligands with glycosphingolipid specificity (P fimbriae or the B subunit of Shiga toxin) were shown to increase the levels of ceramide and to trigger a TLR4-dependent response in epithelial cells. The results show that ceramide activates TLR4 signalling and suggest that this mechanism might allow pathogens to elicit mucosal TLR4 responses by perturbing sphingolipid receptors for virulence ligands like P fimbriae.
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